The first structure–function study of GH151 α‐ l ‐fucosidase uncovers new oligomerization pattern, active site complementation, and selective substrate specificity

Koval'ová T, Kovaľ T, Stránský J, Kolenko P, Dušková J, Švecová L, Vodičková P, Spiwok V, Benešová E, Lipovová P, Dohnálek J, The FEBS Journal (2022) DOI

SASDKZ7 – α-L-Fucosidase isoenzyme 2 from Paenibacillus thiaminolyticus - mutant H503A

Alpha-L-fucosidase H503A
MWexperimental 243 kDa
MWexpected 297 kDa
VPorod 433 nm3
log I(s) 1.12×101 1.12×100 1.12×10-1 1.12×10-2
Alpha-L-fucosidase H503A small angle scattering data  s, nm-1
ln I(s)
Alpha-L-fucosidase H503A Guinier plot ln 1.12×101 Rg: 4.5 nm 0 (4.5 nm)-2 s2
(sRg)2I(s)/I(0)
Alpha-L-fucosidase H503A Kratky plot 1.104 0 √3 sRg
p(r)
Alpha-L-fucosidase H503A pair distance distribution function Rg: 4.7 nm 0 Dmax: 13.5 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Alpha-L-fucosidase H503A PYMOL model
log I(s)
 s, nm-1
Alpha-L-fucosidase H503A DAMMIN model
SAXS data from solutions of α-L-Fucosidase isoenzyme 2 (H503A mutant) in 50 mM potassium phosphate, pH 7.4 were collected using an Anton Paar SAXSpoint 2.0 instrument (Institute of Biotechnology, Czech Academy of Sciences, Vestec, Czech Republic) equipped with an Eiger R 1M detector at a sample-detector distance of 0.8 m and at a wavelength of λ = 0.134 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 2.40 mg/ml was measured at 15°C. 15 successive 60 second frames were collected. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

Storage temperature = UNKNOWN

Alpha-L-fucosidase H503A (Fuk2 H503A)
Mol. type   Protein
Organism   Paenibacillus thiaminolyticus (Bacillus thiaminolyticus)
Olig. state   Tetramer
Mon. MW   74.2 kDa
 
UniProt   K0JCW6 (1-660)
Sequence   FASTA
 
PDB ID   6TVK